Method for obtaining a fluid sample

ABSTRACT

Method for obtaining a fluid sample from a collection device which includes assembling cap and fluid-holding vessel components of the collection device to contain and isolate a specimen retrieval device adjacent a side wall of the vessel. The position of the specimen retrieval device within the collection device is such that the specimen retrieval device does not substantially interfere with the movement of a fluid transfer device into or out of the assembled collection device. The fluid transfer device is used to draw and remove a fluid substance from the collection device for analysis.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of application Ser. No. 10/715,639,filed Nov. 17, 2003, which is a divisional of application Ser. No.09/821,486, filed Mar. 29, 2001, now U.S. Pat. No. 6,806,094, which is acontinuation of application Ser. No. 09/704,210, filed Nov. 1, 2000, nowU.S. Pat. No. 6,716,397, which is a continuation-in-part of applicationSer. No. 09/675,641, filed Sep. 29, 2000, now abandoned, which is acontinuation-in-part of application Ser. No. 09/570,124, filed May 12,2000, now abandoned, which claims the benefit of U.S. ProvisionalApplication No. 60/134,265, filed May 14, 1999, each of whichapplications is hereby incorporated by reference herein.

FIELD OF THE INVENTION

The present invention relates to caps for use in combination withfluid-holding vessels, such as those designed to receive and retainbiological specimens for clinical analysis and patient monitoring ordiagnosis. In particular, the present invention relates to a cap whichis penetrable by a fluid transfer device used to transfer fluids to orfrom a fluid-holding vessel, where the vessel and cap remain physicallyand sealably associated during a fluid transfer.

The present invention further relates to fluid transfer devices whichcan be used to penetrate the caps of the present invention. Inparticular, these fluid transfer devices are adapted to include ribswhich are expected to improve the strength characteristics of the fluidtransfer devices and which may aid in creating passageways for ventingdisplaced air from within a collection device. In addition to or in lieuof these ribs, fluid transfer devices of the present invention mayinclude grooves on their outer surfaces for creating passageways to ventair displaced from the interior of a penetrated collection device. Byproviding means for venting air from within a collection device, fluidtransfer devices of the present invention are expected to exhibitimproved volume accuracy during fluid transfers (e.g., pipetting).

BACKGROUND OF THE INVENTION

Collection devices are a type of cap and vessel combination commonlyused for receiving and storing biological specimens for delivery toclinical laboratories, where the specimens may be analyzed to determinethe existence or state of a particular condition or the presence of aparticular infectious agent. Types of biological specimens commonlycollected and delivered to clinical laboratories for analysis includeblood, urine, sputum, saliva, pus, mucous and cerebrospinal fluid. Sincethese specimen-types may contain pathogenic organisms, it is importantto ensure that collection devices are constructed to be essentiallyleak-proof during transport from the site of collection to the site ofanalysis. This feature of collection devices is particularly critical inthose cases where the clinical laboratory and the collection facilityare remote from one another.

To prevent leakage, collection device caps are typically designed to bescrewed, snapped or otherwise frictionally fitted onto the vesselcomponent, thereby forming an essentially leak-proof seal between thecap and the vessel. In addition to preventing leakage of the specimen,an essentially leak-proof seal formed between the cap and the vessel ofa collection device will also ameliorate exposure of the specimen topotentially contaminating influences from the surrounding environment.This aspect of a leak-proof seal is important for preventing theintroduction of contaminants that could alter the qualitative orquantitative results of an assay.

While a leak-proof seal should prevent specimen seepage duringtransport, the physical removal of the cap from the vessel prior tospecimen analysis presents another opportunity for contamination. Whenremoving the cap, specimen which may have collected on the under-side ofthe cap during transport could come into contact with a practitioner,possibly exposing the practitioner to harmful pathogens present in thefluid sample. And if the specimen is proteinaceous or mucoid in nature,or if the transport medium contains detergents or surfactants, then afilm or bubbles which may have formed around the mouth of the vesselduring transport can burst when the cap is removed from the vessel,thereby disseminating specimen into the environment. It is also possiblethat specimen residue from one collection device, which may havetransferred to the gloved hand of a practitioner, will come into contactwith specimen from another collection device through routine or carelessremoval of the caps. Another risk is the potential for creating acontaminating aerosol when the cap and the vessel are physicallyseparated from one another, possibly leading to false positives orexaggerated results in other specimens being simultaneously orsubsequently assayed in the same general work area throughcross-contamination.

Concerns with cross-contamination are especially acute when the assaybeing performed involves nucleic acid detection and includes anamplification procedure. There are many procedures in use for amplifyingnucleic acids, including the polymerase chain reaction (PCR), (see,e.g., Mullis, “Process for Amplifying, Detecting, and/or Cloning NucleicAcid Sequences,” U.S. Pat. No. 4,683,195), transcription-mediatedamplification (TMA), (see, e.g., Kacian et al., “Nucleic Acid SequenceAmplification Methods,” U.S. Pat. No. 5,399,491), ligase chain reaction(LCR), (see, e.g., Birkenmeyer, “Amplification of Target Nucleic AcidsUsing Gap Filling Ligase Chain Reaction,” U.S. Pat. No. 5,427,930),strand displacement amplification (SDA), (see, e.g., Walker, “StrandDisplacement Amplification,” U.S. Pat. No. 5,455,166), and loop-mediatedisothermal amplification (see, e.g., Notomi et al., “Process forSynthesizing Nucleic Acid,” U.S. Pat. No. 6,410,278). A review ofseveral amplification procedures currently in use, including PCR andTMA, is provided in HELEN H. LEE ET AL., NUCLEIC ACID AMPLIFICATIONTECHNOLOGIES (1997).

Since amplification is intended to enhance assay sensitivity byincreasing the quantity of targeted nucleic acid sequences present in aspecimen, transferring even a minute amount of pathogen-bearing specimenfrom another container, or target nucleic acid from a positive controlsample, to an otherwise negative specimen could result in afalse-positive result. To minimize the potential for creatingcontaminating specimen aerosols, and to limit direct contact betweenspecimens and humans or the environment, it is desirable to have acollection device cap which can be penetrated by a fluid transfer device(e.g., pipette tip) while the cap remains physically and sealablyassociated with the vessel. And, to prevent damage to the fluid transferdevice which could effect its ability to predictably and reliablydispense or draw fluids, the cap design should limit the forcesnecessary for the fluid transfer device to penetrate the cap. Ideally,the collection device could be used in both manual and automated formatsand would be suited for use with pipette tips made of a plasticmaterial.

In addition, when a sealed collection device is penetrated, the volumeof space occupied by a fluid transfer device will displace an equivalentvolume of air from within the collection device. Therefore, it would bedesirable to have a fluid transfer device with means for permitting airto be released from a collection device at a controlled rate as thefluid transfer device penetrates a surface of the collection device(e.g., associated cap). Without such means, a pressurized movement ofair from the collection device into the surrounding environment couldpromote the formation and release of potentially harmful orcontaminating aerosols, or bubbles in those instances where proteins orsurfactants are present in the fluid sample. Therefore, a fluid transferdevice which facilitates a controlled release of air from a penetratedcollection device is needed to prevent or minimize the release of fluidsample in the form of aerosols or bubbles.

SUMMARY OF THE INVENTION

The present invention addresses potential contamination problemsassociated with conventional collection devices by providing anintegrally molded cap which includes an annular flange adapted to gripan inner or outer side wall surface of a vessel at an open end of thevessel, an annular top wall which is substantially perpendicular to theannular flange, an aperture defined by the inner circumference of theannular top wall, and a conical inner wall which tapers inwardly fromthe aperture to an apex located substantially at the longitudinal axisof the cap. The annular flange and the conical inner wall each havesubstantially parallel inner and outer surfaces, and the annular topwall has substantially parallel upper and lower surfaces. (Unlessindicated otherwise, the term “conical,” as used herein with referenceto the inner wall of the cap, shall mean a generally conical shape whichmay be somewhat rounded as the inner wall tapers inwardly from theaperture to the apex.)

In one alternative aspect, the cap of the present invention does notinclude an annular flange adapted to grip a surface of the vessel.Instead, the annular top wall forms an annular ring having a lowersurface which can be affixed to an upper surface of an annular rim ofthe vessel by such means as a fixing agent (e.g., adhesive) or,alternatively, can be integrally molded with the upper surface of thevessel.

In another alternative aspect, the cap of the present invention includesone or more ribs which extend outwardly from the inner surface of theconical inner wall. These ribs can help to form passageways between anouter surface of a fluid transfer device and the inner surface of theconical inner wall of the cap. Furthermore, these ribs will typicallyminimize the surface area of the cap which comes into contact with apenetrating fluid transfer device, thereby limiting frictionalinterference between the fluid transfer device and the cap as the fluidtransfer device is being withdrawn from a penetrated cap.

The present invention addresses potential air displacement problemsassociated with conventional fluid transfer devices penetrating sealedcollection devices by providing a fluid transfer device having a hollowbody which includes one or more ribs extending outwardly from an outersurface, an inner surface, or both the inner and outer surfaces of thefluid transfer device. When the ribs are located on the outer surface,they are expected to facilitate the formation of passageways between theouter surface of the fluid transfer device and a penetrated surfacematerial of a cap. These passageways were found to advantageouslyfacilitate the release of air displaced from a penetrated collectiondevice, while minimizing the formation and/or release of fluid sample inthe form of an aerosol or bubbles. In some cases, the ribs are alsoexpected to improve the strength characteristics of a fluid transferdevice, so that the fluid transfer device (e.g., plastic pipette tips)is less likely to bend or buckle when contacting a penetrable surface.Improved strength characteristics are expected whether the ribs arepositioned on the outer or the inner surface of the fluid transferdevice.

In an alternative aspect, the fluid transfer device of the presentinvention includes one or more grooves recessed from an outer surface ofthe fluid transfer device which can likewise facilitate the formation ofpassageways between the outer surface of the fluid transfer device and apenetrated surface material of a cap. Also contemplated by the presentinvention are fluid transfer devices having both ribs and grooves.

In a first embodiment of the present invention, the conical inner wallhas a single angle with respect to the longitudinal axis of the cap. Thecap of this embodiment is, in a preferred aspect, penetrable by a fluidtransfer device consisting of a plastic pipette tip, and the penetrableportion of the cap does not significantly impair the pipette tip'sability to accurately draw a fluid substance after the cap has beenpenetrated by the pipette tip.

In a second embodiment of the present invention, the conical inner wallof the cap includes a plurality of striations which extend radiallyoutwardly from the apex, or from one or more start-points near the apex,of the conical inner wall. Each of the striations extends partially orfully from the apex, or from a start-point near the apex, of the conicalinner wall to an outer circumference of the conical inner wall. Thestriations may be in the form of grooves, etchings or a series ofperforations on at least one surface of the conical inner wall, and thethickness of each striation is less than the thickness of non-striatedportions of the conical inner wall. The striations were advantageouslyfound to reduce the force needed to penetrate the cap and toconcomitantly create air passageways between portions of the conicalinner wall and the fluid transfer device as sections of conical innerwall, defined by the striations, peeled away from the fluid transferdevice upon penetration.

In a third embodiment of the present invention, the inner surface of theconical inner wall includes one or more ribs which preferably have alongitudinal orientation. The ribs may be elongated structures or, forinstance, protuberances or series of protuberances which aid in formingpassageways for venting displaced air from a penetrated collectiondevice. As indicated above, the ribs should, in some applications,minimize frictional contact between a fluid transfer device and apenetrated surface of a collection device as the fluid transfer deviceis being withdrawn from the penetrated surface.

In a fourth embodiment of the present invention, the annular flange hasan upper portion which extends vertically above the annular top wall, sothat the upper surface of the annular top wall can serve as a ledge forpositioning and maintaining a wick material substantially above theconical inner wall and within the annular flange. The wick may be of anymaterial or combination of materials designed to inhibit the release ofbubbles, aerosols and/or to provide a wiping feature for removing fluidpresent on the outside of a fluid transfer device as it is beingwithdrawn through the cap of a collection device. The wick materialpreferably draws fluid away from the fluid transfer device by means ofcapillary action.

In a fifth embodiment of the present invention, the cap further includesa seal which is affixed to the annular top wall or an annular topsurface of the upper portion of the annular flange, or is otherwisefixedly positioned within an inner surface of the annular flange (e.g.,a hollow-centered resin disk with a seal affixed thereto and sized tofrictionally fit within an inner surface of the annular flange and topermit passage therethrough by a fluid transfer device). While the sealis preferably penetrable with a fluid transfer device, the seal may beapplied to or associated with the cap in such a way that it can beseparated from the cap prior to penetration with a fluid transferdevice. The seal may be provided to protect the conical inner wall (andthe wick, if present) from contaminants, to limit the release of anaerosol from the collection device once an associated cap has beenpenetrated and/or to retain the wick within the annular flange. Asindicated, the seal is preferably made of a penetrable material, such asa metallic foil or plastic, and is affixed to the cap so that itcompletely or partially covers the conical aperture prior topenetration.

In a sixth embodiment of the present invention, a cap is provided whichcan be penetrated by a plastic pipette tip by applying a force of lessthan about 8 pounds force (35.59 N) to a surface of the cap. The cap ofthis embodiment preferably includes a wick positioned above or below apenetrable surface material of the cap and requires less than about 4pounds force (17.79 N) pressure for the pipette tip to penetrate. Whenincluded, the wick is arranged in the cap so that it can at leastpartially arrest the movement of an aerosol or bubbles from anassociated vessel during and/or after penetration of the cap by theplastic pipette tip.

In a seventh embodiment of the present invention, an overcap containinga wick is provided which can be positioned over a cap of the presentinvention. An annular top wall of the overcap includes an innercircumference which defines an aperture which has been sized to receivea fluid transfer device for penetrating the conical inner wall of thecap. Ribs may be further included on an inner surface of an annularflange of the overcap to provide a frictional fit between the innersurface of the overcap and the annular outer flange of the cap. A sealmay also be applied to the annular top wall of the overcap to furtherminimize aerosol or bubble release from a collection device once the caphas been penetrated and/or to retain the wick within the annular flangeof the overcap. The overcap, which provides the benefits of aerosol andbubble containment in a separate component, may be optionally employed,for example, with a collection device having a cap lacking a wick whenthe sample to be removed and analyzed is suspected of containing atarget nucleic acid analyte which is to be amplified before a detectionstep is performed.

In an eighth embodiment of the present invention, a fluid transferdevice is provided which may be used to facilitate penetration of thecap or overcap of the present invention and/or which may improve ventingof air displaced from an enclosed collection device as it is beingentered by the fluid transfer device. This particular fluid transferdevice is hollow in construction (although the fluid transfer device maybe outfitted with an aerosol impeding filter), designed to be engaged bya probe or extension associated with a robotic or manually operatedfluid transfer apparatus for drawing and/or dispensing fluids, andincludes one or more ribs. These ribs extend outward from an outersurface of the body of the fluid transfer device and preferably have alongitudinal orientation starting from a point or points at or near thedistal end of the fluid transfer device. (As used herein, the term“longitudinal orientation” shall mean a generally lengthwiseorientation.)

In a ninth embodiment of the present invention, a plastic pipette tip isprovided which has hollow tubular and conical sections for the passageof air and/or fluids therethrough and one or more lower ribs located onthe conical section which extend outward from an outer surface of theconical section. These lower ribs are expected to provide the samebenefits attributable to the eighth embodiment of the present invention.

In a tenth embodiment of the present invention, a plastic pipette tip isprovided which has hollow tubular and conical sections for the passageof air and/or fluids therethrough and one or more lower ribs located onthe conical section which extend inward from an inner surface of theconical section. As with the eighth embodiment, these lower ribs areexpected to facilitate penetration of the caps and overcap of thepresent invention

In an eleventh embodiment of the present invention, a plastic pipettetip is provided which has hollow tubular and conical sections for thepassage of air and/or fluids therethrough and one or more upper ribs onthe tubular section which extend outward from an outer surface of thetubular section, with at least one of these upper ribs having a terminusat or near the distal end of the tubular section. These upper ribs aredesigned to aid in the formation of air gaps or passageways between thepenetrated surface material of a cap and the pipette tip to facilitatethe movement of air displaced from the interior of a collection deviceas it is being entered by the pipette tip and/or so that the airpressures inside and outside of the collection device can quicklyequilibrate upon penetration of the cap.

In a twelfth embodiment of the present invention, a plastic pipette tipis provided which combines the lower and upper ribs of the ninth andeleventh or tenth and eleventh embodiments described above, where thelower ribs may be distinct from the upper ribs or pairs of lower andupper ribs may form continuous ribs extending from a point or points onthe conical section to a point or points on the tubular section.

In a thirteenth embodiment of the present invention, a fluid transferdevice is provided which may be used to improve venting of air displacedfrom an enclosed collection device as it is being penetrated by thefluid transfer device. This fluid transfer device is hollow inconstruction, designed to be engaged by a probe or extension associatedwith a robotic or manually operated fluid transfer apparatus for drawingand/or dispensing fluids, and includes one or more grooves. Thesegrooves are recessed from an outer surface of the body of the fluidtransfer device and preferably have a longitudinal orientation. Thegrooves of this embodiment may be used alone or in combination with theribs of any one of the eighth, ninth, tenth, eleventh and twelfthembodiments described above.

In a fourteenth embodiment of the present invention, a method isprovided for displacing air from a collection device having an enclosedchamber. In this method, a surface of the collection device ispenetrated with a fluid transfer device and air is released from thecollection device through a passageway formed between the surface of thecollection device and an outer surface of the fluid transfer device. Thefluid transfer device used in this method could be the fluid transferdevice of the thirteenth embodiment described above.

In a fifteenth embodiment of the present invention, another method isprovided for displacing air from a collection device having an enclosedchamber. In this method, a surface of the collection device ispenetrated with a fluid transfer device and air is released from thecollection device through a passageway formed adjacent to a point ofcontact between the surface of the collection device and a ribpositioned on an outer surface of the fluid transfer device. The fluidtransfer device used in this method could be the fluid transfer deviceof any one of the eighth, ninth, twelfth and thirteenth embodimentsdescribed above.

In a sixteenth embodiment of the present invention, a method is providedfor removing a fluid substance from a collection device which includespenetrating a cap component of the collection device with a plasticfluid transfer device by applying a force of less than about 8 poundsforce (35.59 N) to a surface of the cap. Once the cap has beenpenetrated, a fluid substance present in a vessel component of thecollection device is withdrawn by the fluid transfer device beforeremoving the fluid transfer device from the collection device.

In a seventeenth embodiment of the present invention, another method isprovided for removing a fluid substance from a collection device whichincludes piercing a surface of the collection device after contactingthe surface of the collection device or a surface of the fluid transferdevice with a lubricant, such as a detergent. Subsequent to piercing thesurface of the collection device, the fluid transfer device draws atleast a portion of a fluid substance contained in a vessel component ofthe collection device before being completely removed from thecollection device. The lubricant, which may be contained in aspecimen-bearing transport medium held by the vessel, is expected toreduce the frictional forces between the surface of the collectiondevice and the outer surface of the fluid transfer device as the fluidtransfer device is being removed from the collection device.

In an eighteenth embodiment of the present invention, yet another methodis provided for removing a fluid substance from a collection devicewhich includes a first step for puncturing a surface of the collectiondevice with a fluid transfer device followed by a second step forpenetrating or entering the collection device so that a distal end ofthe fluid transfer device comes into contact with a fluid substancecontained in a vessel component of the collection device. The first andsecond steps of this method may be performed at the same or differentspeeds. When the steps are performed at the same speed, a pauseinterrupts the movement of the fluid transfer device between the firstand second steps. And when the steps are performed at different speeds,the speed of the fluid transfer device in the second step is greaterthan the speed of the fluid transfer device in the first step. Anintervening pause may also be introduced between the first and secondsteps when these steps are carried out at different speeds. Aftercontacting the fluid substance, the fluid transfer device draws at leasta portion of the fluid substance before it is completely removed fromthe collection device. This two-step penetration method was found toimprove the volume accuracy of fluid samples being withdrawn fromcollection devices.

In a nineteenth embodiment of the present invention, a further method isprovided for removing a fluid substance from a collection device whichincludes penetrating a surface of a collection device with aconically-shaped pipette tip and then inserting the pipette tip into thecollection device until a distal end of the pipette tip comes intocontact with the fluid substance. After contacting the fluid substance,the distal end of the pipette tip is partially or fully removed from thefluid substance a sufficient distance so that one or more passagewaysare formed or enlarged between an outer surface of the pipette tip andthe penetrated surface of the collection device. (The passageways aid inventing of air from within the collection device, facilitating greatervolume accuracy during fluid aspirations.) The pipette tip then draws atleast a portion of the fluid substance contained in the collectiondevice before the pipette tip is completely removed from the collectiondevice.

In a twentieth embodiment of the present invention, yet a further methodis provided for removing a fluid substance from a collection devicewhich includes positioning a specimen retrieval device (e.g., swab)along an inner surface of a side wall of a vessel component of thecollection device by means of fixedly associating the vessel with a capcomponent of the collection device. The cap is then penetrated with afluid transfer device which draws a fluid substance from the vesselbefore the fluid transfer device is removed from the collection device.

In a twenty-first embodiment of the present invention, a method isprovided for containing an aerosol substantially inside of a collectiondevice after a cap associated with the collection device has beenpenetrated by a fluid transfer device, such as a plastic pipette tip,where the cap contains a wick. Penetration of the cap results in theformation of at least one passageway which may be partially open duringpenetration of the cap by the fluid transfer device and/or duringremoval of the fluid transfer device from the collection device. Thewick, therefore, may aid in containing an aerosol within the collectiondevice (either partially or completely) as the fluid transfer device isentering an interior chamber of the collection device, as the fluidtransfer device is being withdrawn from the collection device and/orafter the fluid transfer device has been completely withdrawn from thecollection device. The material selected for the wick, and itsarrangement inside of the cap, should be such that the material will notsubstantially impede movement of the fluid transfer device into or outof the collection device. This method is particularly useful when thecollection device contains a fluid sample suspected of having a targetnucleic acid analyte which will be subsequently amplified using anyknown amplification procedure prior to a detection step.

Caps of the present invention may be provided in packaged combinationwith at least one of a vessel, a reagent (e.g., transport medium orpositive control), an overcap, a fluid transfer device and a specimenretrieval device (e.g., swab or other type of probe used for specimencollection). Likewise, the overcaps of the present invention may beprovided in packaged combination with at least one of a cap, a vessel, areagent, a fluid transfer device, and a specimen retrieval device. To bein packaged combination, it is to be understood that the recited itemsmerely need to be provided in the same container (e.g., mail or deliverycontainer for shipping), and it is not a requirement that the items beper se physically associated with one another in the container orcombined in the same wrapper.

These and other features, aspects, and advantages of the presentinvention will become apparent to those skilled in the art afterconsidering the following detailed description, appended claims andaccompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows an exploded perspective view of a collection deviceaccording to the present invention.

FIG. 2 shows an enlarged top plan view of a cap component of thecollection device illustrated in FIG. 1.

FIG. 3 shows an enlarged bottom view of the cap illustrated in FIG. 1.

FIG. 4 shows an enlarged top plan view of another cap embodiment of thepresent invention.

FIG. 5 shows an enlarged partial section side view of the collectiondevice illustrated in FIG. 1, taken along the 5-5 line thereof.

FIG. 6 shows an enlarged partial section side view of another collectiondevice according to the present invention.

FIG. 7 shows the enlarged partial section side view of the collectiondevice illustrated in FIG. 5, where the collection device has beenpenetrated by a fluid transfer device and contains an immobilizedspecimen retrieval device.

FIG. 8 shows an enlarged top plan view of the cap illustrated in FIG. 5after the fluid transfer device has been removed therefrom.

FIG. 9 shows an enlarged partial section side view of an overcap andcollection device combination according to the present invention.

FIG. 10 shows an enlarged side elevation view of a pipette tip accordingto the present invention.

FIG. 11 shows another enlarged side elevation view of the pipette tipillustrated in FIG. 10.

FIG. 12 shows an enlarged perspective view of a distal end portion ofthe pipette tip illustrated in FIG. 10.

FIG. 13 shows an enlarged bottom section view of the pipette tipillustrated in FIG. 11, taken along the 13-13 line thereof.

FIG. 14 shows an enlarged side elevation view of another pipette tipaccording to the present invention.

FIG. 15 shows another enlarged side elevation view of the pipette tipillustrated in FIG. 14.

FIG. 16 shows an enlarged perspective view of a distal end portion ofthe pipette tip illustrated in FIG. 14.

FIG. 17 shows an enlarged side elevation view of another pipette tipaccording to the present invention.

FIG. 18 shows an enlarged side section view of the pipette tipillustrated in FIG. 17, taken along the 17-17 line thereof.

FIG. 19 shows an enlarged bottom section view of the pipette tipillustrated in FIG. 17, taken along the 19-19 line thereof.

FIG. 20 shows an enlarged side elevation view of another pipette tipaccording to the present invention.

FIG. 21 shows an enlarged side elevation view of another pipette tipaccording to the present invention.

FIG. 22 shows an enlarged side elevation view of another pipette tipaccording to the present invention.

FIG. 23 shows another enlarged side elevation view of the pipette tipillustrated in FIG. 22.

FIG. 24 shows an enlarged bottom section view of the pipette tipillustrated in FIG. 23, taken along the 24-24 line thereof.

FIG. 25 shows an enlarged bottom section view of the pipette tipillustrated in FIG. 23, taken along the 25-25 line thereof.

FIG. 26 shows an enlarged top plan view of the pipette tip illustratedin FIG. 15 in cross-section, taken along the 26-26 line thereof.

FIG. 27 shows an enlarged top plan view of the pipette tip illustratedin FIG. 23 in cross-section, taken along the 27-27 line thereof.

FIG. 28 shows an enlarged top plan view of another cap according to thepresent invention.

FIG. 29 shows an enlarged section side view of the cap illustrated inFIG. 28, taken along the 29-29 line thereof.

FIG. 30 shows an enlarged top plan view of the cap illustrated in FIG.28, after the cap has been penetrated by a fluid transfer device shownin cross-section.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

With reference to the figures, the cap 20A-C of the present inventioncan be combined with a vessel 50 to receive and store fluid specimensfor subsequent analysis, including analysis with nucleic acid-basedassays or immunoassays diagnostic for a particular pathogenic organism.When the desired specimen is a biological fluid, the specimen can be,for example, blood, urine, saliva, sputum, mucous or other bodilysecretion, pus, amniotic fluid, cerebrospinal fluid or seminal fluid.However, the present invention also contemplates materials other thanthese specific biological fluids, including, but not limited to, water,chemicals and assay reagents, as well as solid substances which can bedissolved in whole or in part in a fluid milieu (e.g., tissue specimens,stool, environmental samples, food products, powders, particles andgranules). Vessels 50 used with the cap 20A-C of the present inventionare preferably capable of forming a substantially leak-proof seal withthe cap 20A-C and can be of any shape or composition, provided thevessel 50 is shaped to receive and retain the material of interest(e.g., fluid specimen or assay reagents). Where the vessel 50 contains aspecimen to be assayed, it is important that the composition of thevessel 50 be essentially inert so that it does not significantlyinterfere with the performance or results of an assay.

The cap 20A-C of the present invention may be prepared from a number ofdifferent polymer and heteropolymer resins, including, but not limitedto, polyolefins (e.g., high density polyethylene (“HDPE”), low densitypolyethylene (“LDPE”), a mixture of HDPE and LDPE, or polypropylene),polystyrene, high impact polystyrene and polycarbonate. An example of anHDPE is sold under the tradename Alathon M5370 and is available fromPolymerland of Huntsville, N.C.; an example of an LDPE is sold under thetradename 722 and is available from The Dow Chemical Company of Midland,Mich.; and an example of a polypropylene is sold under the tradenameRexene 13T10ACS279 and is available from the Huntsman Corporation ofSalt Lake City, Utah. Although LDPE is a softer, more malleable materialthan HDPE, the softness of LDPE creates more frictional resistance whena threaded cap is screwed onto a threaded vessel than when a cap isformed of the more rigid HDPE material. And, while a cap made of HDPE ismore rigid than one made of LDPE, this rigidity tends to make an HDPEcap more difficult to penetrate than one made of LDPE. Although the cap20A-C of the present invention is preferably comprised of HDPE, it canalso be comprised of a combination of resins, including, for example, amixture of LDPE and HDPE, preferably in a mixture range of about 20%LDPE:80% HDPE to about 50% LDPE:50% HDPE by volume.

Based on the guidance provided herein, those skilled in the will be ableto select a resin or mixture of resins having hardness and penetrationcharacteristics which are suitable for a particular application, withouthaving to engage in anything more than routine experimentation.Additionally, skilled artisans will realize that the range of acceptablecap 20A-C resins will also depend on the nature of the resin used toform the vessel 50, since the properties of the resins used to formthese two components will affect how well the cap and vessel of thecollection device 10 can form a leak proof seal and the ease with whichthe cap can be securely screwed onto the vessel. (Polypropylene iscurrently the material of choice for the vessel 50.) To modify therigidity and penetrability of a cap, those skilled in the art willappreciate that the molded material may be treated, for example, byheating, irradiating or quenching.

Regardless of the type or mixture of resins chosen, the cap 20A-C ispreferably injection molded as a unitary piece using procedureswell-known to those skilled in the art of injection molding, including amulti-gate process for facilitating uniform resin flow into the capcavity used to form the shape of the cap. Uniform resin flow isdesirable for achieving consistency in thickness, which is especiallyimportant for the penetrable surface of the cap 20A-C. After preparingthe integrally molded cap 20A-C, a wick 90 may be provided within theaperture defined either by an inner circumference 25 of the annular topwall 22, (see FIG. 2), or by the circumference of an inner surface 123of the upper portion 46 of the annular outer flange 40A (see FIG. 6).The wick 90 is preferably positioned above the conical inner wall 33 ofthe cap 20A-C to aid in further containing and limiting thedissemination of an aerosol outside of the collection device 10. Inaddition, a seal 80 may be applied to an upper surface 24 of an annulartop wall 22 (cap 20A-B) or an annular top surface 48 (cap 20C) toprovide a protective cover over the aperture above the conical innerwall 33 of the cap (and to retain the wick 90, if present, in the cap),as depicted in FIGS. 5 and 6.

While the outer circumference 38 of the conical inner wall 33 maycoincide with the inner circumference 25 of the annular top wall 22 in asingle plane (not shown), such that there is no annular inner flange,the cap 20A of FIG. 5 is a preferred embodiment since it includes anannular inner flange 49 which extends substantially vertically from theouter circumference 38 of the conical inner wall 33 to the innercircumference 25 of the annular top wall 22, providing the additionalvertical space in the aperture required for receiving a wick 90.However, when a wick 90 is to be included in the cap 20A-C, an extensionof the annular outer flange 40A, as illustrated in FIG. 6, isparticularly preferred. In this arrangement, the annular outer flange40A has an upper portion 46 located above the upper surface 24A of theannular top wall 22A, and is constructed so that an inner surface 123 ofthe upper portion 46 of the annular outer flange 40A terminates at theupper surface 24A of the annular top wall 22A. With this preferredarrangement, the inner circumference 25 of the annular top wall 22A issmaller than the circumference defined by the inner surface 123 of theupper portion 46 of the annular outer flange 40A. In this way, the uppersurface 24A of the annular top wall 22A can function as a ledge forpositioning and maintaining a wick 90 above the conical inner wall 33.

Inclusion of a wick 90 not only helps to retard the movement of anaerosol from the vessel 50 to the environment, it can also beconstructed to perform a wiping action on the outside of a fluidtransfer device as the fluid transfer device is being removed from thevessel 50 and cap 20A-C. In a preferred mode, the wick 90 functions todraw fluids away from the outside of the fluid transfer device by meansof capillary action. As used herein, however, the term “wick” refers toa material which performs a wiping function to remove fluids present onthe outside of a fluid transfer device and/or an absorbing function tohold fluids removed from the outside of a fluid transfer device.Examples of wick 90 materials which may be used with the cap 20A-C ofthe present invention include, but are not limited to, pile fabrics,sponges, foams (with or without a surface skin), felts, sliver knits,GORE-TEX® fabrics, spandex, and other materials, both natural andsynthetic. These materials may also be mechanically or chemicallytreated to further improve the intended functions of the wick 90. Forexample, napping may be used to increase the surface area and,therefore, the fluid holding capacity of a wick 90. The material of thewick 90 might also be pre-treated with a wetting agent, such as asurfactant, to lower the surface tension of a fluid present on an outersurface of a fluid transfer device. An acrylic binder might be used, forexample, to actually bind the wetting agent to the wick 90 material.

If the fluid transfer device does not have a uniform diameter, as is thecase with most standard air displacement pipette tips, then the wick 90is preferably made of a resilient material whose original shape isrestored or substantially restored as the fluid transfer device is beingremoved from the collection device 10. Thus, materials such as pilefabric, sponges, foams and spandex are preferred because of theirability to rebound rapidly after exposure to compressive forces. Pilefabric is a particularly preferred wick 90 material, an example of whichincludes a {fraction (3/8)} inch (9.53 mm) pile fabric of acrylicconstruction which is available from Roller Fabrics of Milwaukee, Wis.as Part No. ASW112. Other acceptable pile fabrics are made of acrylicand polyester materials, range in size from {fraction (1/4)} inches(6.35 mm) to {fraction (5/16)} inches (7.95 mm) and are available fromMount Vernon Mills, Inc. of LeFrance, S.C. as Part Nos. 0446, 0439 and0433. The wick 90 material is preferably inert with respect to a fluidsample contained within the vessel 50.

Because wick 90 materials are designed to draw fluids away from theexterior of fluid transfer devices and/or to capture fluids in the formof an aerosol and/or bubbles, the material and dimensions of the wickmust be chosen to avoid excessive saturation with fluid. If the wick 90becomes overly saturated, fluid may not be adequately wiped from theexterior of the fluid transfer device and/or bubbles may be producedupon insertion of the fluid transfer device and/or displacement of airfrom within the collection device 10. Thus, it is important to adapt thesize and adsorptive properties of the wick 90 in order to achieveadequate wiping and aerosol and/or bubble containment for a given cap20A-C configuration, fluid transfer device and fluid substance, giventhe number of anticipated fluid transfers the wick will be exposed to.Hence, as the volume of liquid that the wick 90 will be exposed to in anapplication increases, the amount of wick material and/or its absorptiveproperties may need to be adjusted so that the wick does not becomeoverly saturated during use.

It is also important that the wick 90 be constructed and arranged in thecap 20A-C so that the flow of air out of the collection device 10 isrelatively unimpeded. While this property is important when the wick 90is dry, it is especially important when the wick has absorbed themaximum volume of fluid expected for a given application. However, itshould be recognized that this property of the wick 90 needs to bebalanced with the requirement that the wick have sufficient density totrap an escaping aerosol and/or bubbles. Therefore, those skilled in theart will need to select or design wick 90 materials having matrices thatare capable of trapping an aerosol and bubbles, while simultaneouslypermitting air to be vented from the collection device 10 once theunderlying surface material of the penetrable cap 20A-C has beenpierced.

As shown in FIG. 6, the wick 90 is preferably sized to fit beneath thehorizontal plane of the annular top surface 48 of the cap 20C (or theupper surface 24 of the annular top wall 22 of the cap 20A-B) and abovethe annular top wall 22A, where it is restrained by the seal 80 andannular top wall 22A. To better ensure that the wick 90 is notsubstantially moved from this location by frictional contact with afluid transfer device penetrating or being removed from the cap 20A-C,at least one annular shelf (not shown) above or below the wick andextending inwardly from an inner surface 21, 123 of the cap could beprovided. Such an annular shelf would be particularly advantageous wherethe cap 20A-C does not include a seal 80. Moreover, in an effort tofurther impede the mobility of the wick 90, the wick could be glued orotherwise adhered to at least one of the suggested annular shelves, theseal 80 and the annular top wall 22A. Alternatively, the wick 90 may beglued or otherwise adhered to the inner surface 123 of the upper portion46 of the annular outer flange 40A.

In a preferred embodiment, the aperture defined by the inner surface 123of the upper portion 46 of the annular outer flange 40A is sealed with ametallic foil 80 (or foil laminate) using, for example, a pressuresensitive adhesive which is applied to the annular top surface 48 (cap20C) or the upper surface 24 of the annular top wall 22 (cap 20A-B). Thematerial and configuration of the wick 90 should be such that it createsminimal frictional interference with the fluid transfer device when itis inserted into or withdrawn from the cap and vessel 50. In the case ofa sponge or foam, for example, this may require boring a hole orcreating one or more slits in the center of the wick 90 which are sizedto minimize frictional interference but, at the same time, to providesome frictional interference with the fluid transfer device so thataerosol transmission is limited and the wiping action is performed. If apile fabric is employed as the wick 90, the pile fabric is preferablyarranged so that the free ends of individual fibers are oriented inwardtoward a longitudinal axis 30 of the cap 20A-C and away from the pilefabric backing which is arranged in the cap in a generally circularfashion within an inner surface 21 of the annular inner flange 49 or theinner surface 123 of the upper portion 46 of the annular outer flange40A. Care should be taken not to wind the pile fabric so tightly that itwill create excessive frictional interference with a fluid transferdevice penetrating the cap 20A-C, thereby substantially impedingmovement of the fluid transfer device. The movement of a fluid transferdevice is deemed “substantially impeded” if the force required topenetrate the wick 90 is greater than the force required to penetratethe cap which contains it. The force required to penetrate the wick 90is preferably less than about 4.0 pounds force (17.79 N), morepreferably less than about 2.0 pounds force (8.90 N), even morepreferably less than about 1.0 pound force (4.45 N), and most preferablyless than about 0.5 pounds force (2.22 N). A method and instrumentationwhich can be used to determine the force required to penetrate a wick 90material is described in the Example infra.

When the seal 80 is included, it is preferably made of a plastic film(e.g., biaxial polypropylene) or metallic foil material (e.g., aluminumfoil), which can be affixed to the annular top surface 48 (cap 20C) orthe upper surface 24 of the annular top wall 22 (cap 20A-B) using meanswell known to those skilled in the art, including adhesives. A metallicseal 80 may further include a plastic liner, such as a thin veneer ofHDPE applied to one or both surfaces of the metallic material, whichpromotes attachment of the seal to the annular top wall 22 when a heatinduction sealer is used. Heat induction sealing is a well known processand involves the generation of heat and the application of pressure tothe surface being sealed, which, in this case, is the annular topsurface 48 (cap 20C) or the upper surface 24 of the annular top wall 22(cap 20A-B). The heat is used to soften the material of the annular topsurface 48 or the annular top wall 22 (and the seal 80 if it includes aresin veneer) for permanently receiving the seal 80, and pressure isapplied to the cap 20A-C while the seal becomes affixed to the annulartop surface 48 or the upper surface 24 of the annular top wall 22. Anyknown ultrasonic welding procedure using either high frequency or highamplitude sound waves may also be used to affix the seal 80 to the cap20A-C.

Where aerosol release from the collection device 10 is a particularconcern, the seal 80 may be used to further reduce the amount of aerosolwhich can be released from the collection device when the conical innerwall 33 of the cap 20A-C is penetrated. Under these circumstances, thematerial selected for the seal 80 should experience minimal tearing whenthe fluid transfer device, such as a pipette tip or fluid-transportingneedle or probe, passes through it. Some tearing, however, is desirableto avoid creating a vacuum within the collection device 10 once the cap20A-C has been penetrated. An example of a pipette that can be used withthe cap 20A-C of the present invention is a Genesis series 1000 μlTecan-Tip (with filter), available from Eppendorf-Netherler-Hinz GmbH ofHamburg, Germany. In addition to limiting the amount of aerosol releasedfrom the collection device 10, the seal 80 can also serve to protect theconical inner wall 33 of the cap 20A-C and/or the inserted wick 90 fromundesirable environmental contaminants.

As exemplified in FIG. 5, the cap 20A-C of the present invention isdesigned to include a conical inner wall 33 which tapers inwardly fromthe aperture which is defined by the inner circumference 25 of theannular top wall 22, (see FIG. 2), to an apex 34 located substantiallyat the longitudinal axis 30 of the cap. (The apex 34 may have a roundedor concave configuration and need not have the pointed shape shown inthe figures.) The shape of the conical inner wall 33 aids in guiding thefluid transfer device to the apex 34 in the conical inner wall 33 wherethe fluid transfer device 70 will penetrate the cap 20A-C, as shown inFIG. 7. Therefore, the angle of the conical inner wall 33 should bechosen so that penetration of the apex 34 by the tip 71 of the fluidtransfer device 70 is not substantially impeded. Thus, the angle of theconical inner wall 33, with respect to the longitudinal axis 30, ispreferably about 25° to about 65°, more preferably about 35° to about55°, and most preferably about 45°±5°. Ideally, the conical inner wall33 has a single angle with respect to the longitudinal axis 30.

As shown in FIG. 7, it was discovered that the shape of the conicalinner wall 33 of the cap 20A-C of the present invention can alsofunction to position a specimen retrieval device, such as aspecimen-bearing swab 130 or other type of probe, along an inner surface59 of a side wall 58 of the vessel 50 so that it does not significantlyinterfere with the movement of a fluid transfer device either into orout of the collection device 10. To ensure that the swab 130 issufficiently isolated from the pathway of the fluid transfer devicewithin the collection device 10, the swab 130 will need to be sized sothat it fits snugly beneath an outer surface 37 of the conical innerwall 33 and along the inner surface 59 of the side wall 58 of the vessel50, (see FIG. 7), when the collection device is fully assembled. One wayto achieve this snug fit is to use a swab 130 which has beenmanufactured to include a mid-section score line (not shown), therebypermitting an upper portion of the swab 130 to be manually snapped-offand discarded after use, leaving only the specimen-bearing, lowerportion of the swab in the collection device 10. The precise location ofthe score line on the swab 130 will need to be determined based upon theinterior dimensions of the collection device 10 when the cap 20A-C isfrictionally-fitted onto the vessel 50. Breakable swabs are fullydescribed in U.S. Pat. No. 5,623,942, the contents of which are herebyincorporated by reference herein.

Another embodiment of the present invention is depicted in FIG. 9 andincludes an overcap 100, preferably constructed of an injected moldedplastic which has been adapted to fit over the cap 20A-B shown in FIGS.2-5 (generally without the seal 80), preferably forming a frictional fitbetween the annular outer flange 40 of the cap 20 and a portion of aninner surface 101 of the annular flange 102 of the overcap. To achievethis frictional fit between the cap 20A-B and the overcap 100, theovercap may be configured to include one or more ribs 103 which extendinwardly from the inner surface 101 of the overcap and which physicallycontact with the annular outer flange 40 when the overcap is positionedover the cap. The overcap 100 of this embodiment contains a wick 90which is fixedly positioned within the inner surface 101 of the annularflange 102 and beneath a lower surface 105 of an annular top wall 104 ofthe overcap by means of, for example, a frictional fit or adhesive. Thewick 90 can be used for any of the reasons discussed hereinabove and maybe made of any material having the aerosol retarding or wipingproperties referred to supra. A seal 80 may also be included, forinstance, to act as an additional barrier to the flow of an aerosol fromthe collection device 10 when the conical inner wall 33 is penetrated bya fluid transfer device. When used, the seal 80 is preferably applied tothe annular top wall 104 of the overcap 100 using conventional methods,including the heat induction and ultrasound methods discussedhereinabove. To permit penetration of the conical inner wall 33 of thecap 20A-B by a fluid transfer device, the annular top wall 104 of theovercap 100 includes an aperture 107 sized to receive the fluid transferdevice, where the size of the aperture 107 is large enough so that theannular top wall 104 does not interfere with the movement of the fluidtransfer device into and out of the vessel 50 component of thecollection device 10.

Included in the conical inner wall 33 of the preferred cap 20A-C are aplurality of striations 35 which extend radially outwardly from the apex34, or from one or more start-points 31 near the apex, (see, e.g., FIG.4), toward the outer circumference 38 of the conical inner wall 33. (Toavoid cluttering FIGS. 2-6 and 8, those skilled in the art willappreciate that only some of the multiple start-points, end-points 27,striations 35 and pie-shaped sections 26 which are clearly illustratedin these drawings are identified with reference numerals.) Where astriation 35 extends from a start-point 31 “near” the apex 34, thestart-point 31 is located on the conical inner wall 33 within a distanceof at least about 0.05 inches (1.27 mm) from the apex 34, and preferablywithin a distance of at least about 0.025 inches (0.635 mm) from theapex 34. When the start-points 31 of the striations 35 in the conicalwall 33 are all positioned slightly away from the apex 34, it wasdiscovered that a more uniform resin thickness in the apex 34 could beachieved during the injection molding process and that the striations 35tended to “open” more evenly upon penetration, as described infra.

The striations 35, as shown in FIGS. 1-6, 8 and 9, were discovered toenhance penetration of the conical inner wall 33 by a fluid transferdevice. Examples of striations 35 in the conical inner wall 33 of thecap 20A-C include grooves, etchings or a series of perforations whichcan be formed on a core pin using known injection molding techniques orwhich can be physically “etched” or pierced with a cutting toolfollowing formation of the cap using well known techniques. Thestriations 35 may be of any number sufficient to improve penetrabilityof the conical inner wall 33 of the cap 20A-C, as determined by areduction in the force required to penetrate the cap. Notwithstanding,the number of striations 35 on a cap 20A-C is preferably from about 3 toabout 12, more preferably from about 6 to about 10, and most preferablyabout 8. In one embodiment shown in FIG. 2, the striations 35 all extendan approximately equal distance from the apex 34 to form generallywedge-shaped sections 26 on the conical inner wall 33 when an imaginaryline 28 is circumferentially drawn to connect the end-points 27 of thestriations 35. A similar configuration is shown for the fully extendedstriations 35 in FIG. 4. These wedge-shaped sections 26 illustrated inFIGS. 2 and 4 are preferably of the same approximate size and shape. Thestriations 35 may be formed on either the inner surface 36 of theconical inner wall 33 or the outer surface 37 of the conical inner wall33 or both surfaces 36, 37.

When striations 35 are included with a cap 20A-C of the presentinvention, the force needed to penetrate the cap with a fluid transferdevice is less than the force needed to penetrate a cap of the samematerial, shape and dimensions, but which includes no striations 35.Preferably, the force required to penetrate a cap 20A-C having aplurality of striations 35 is no more than about 95% of the forcerequired to penetrate a cap of identical material, shape and dimensionsbut which has no striations 35. (To “penetrate” a cap 20A-C, a fluidtransfer device need only pierce the conical inner wall 33, preferablyat or near the apex 34.) This percentage is more preferably no more thanabout 85%, even more preferably no more than about 75%, and mostpreferably no more than about 65%. When the fluid transfer device 70includes a beveled tip 71, as shown in FIG. 7, this percentage isideally no more than about 50%. For all caps of the present invention,whether striated or unstriated, the preferred force needed by a plasticfluid transfer device (i.e., pipette tip) to penetrate the cap is lessthan about 8.0 pounds force (35.59 N), more preferably less than about6.0 pounds force (26.69 N), and most preferably less than about 4.0pounds force (17.79 N). The force needed to penetrate a cap can bedetermined using the equipment, materials and protocol described in theExample infra.

A particularly preferred fluid transfer device for use with the cap20A-C of the present invention is a pipette tip 70A-C shown in FIGS.10-19. This pipette tip 70A-C includes one or more lower ribs 151A-C,152A-C which are preferably, although not necessarily, longitudinal inorientation and extend outward from an outer surface 153 at the distalend of the pipette tip 70A-B or inward from an inner surface 157 at thedistal end of the pipette tip 70C. (Also contemplated by the term“ribs”, as applied to any embodiment herein, is a series of abbreviatedor interrupted ribs (not shown) which, for example, may be in the formof a series of protuberances which are the same or different in size andshape and which are equally or unequally spaced apart.) The addition ofthese lower ribs 151A-C, 152A-C was found to strengthen the pipette tip70A-C so that it can more easily penetrate the cap 20A-C withoutbending. Bending of the pipette tip 70A-C could prevent penetration ofthe cap 20A-C, occlude an orifice 161 of the pipette tip and/ormisdirect a fluid stream subsequently dispensed from the pipette tip.

While the lower ribs 151A-B, 152A-B preferably have a longitudinalorientation on the outer surface 153 of the pipette tip 70A-B, it isusually desirable to have at least one lower rib structure 151Apositioned on the outer surface 153 at the distal end of the pipette tip70A so that a terminus 162A of the lower rib structure 151Aco-terminates with the point 155A of a beveled tip 71A. (It is notedthat lower ribs 151A-C, 152A-C can also be used with pipette tips whichhave a flat or blunt-ended surface surrounding the orifice 161 at thedistal end (not shown).) If the pipette tip 70A-B includes more than onelower rib structure, then the lower ribs 151A-B, 152A-B are preferablycircumferentially spaced-apart at equal distances on the outer surface153 at the distal end of the pipette tip 70A-B, although this precisearrangement of lower ribs 151A-B, 152A-B is not a requirement.

Ideally, the pipette tip 70A-C is a conventional single-piece, plasticpipette tip modified to include the lower ribs 151A-C, 152A-C duringmanufacture using any well-known injection molding procedure. An exampleof acceptable pipette tip, prior to any of the modifications describedherein, is an ART® 1000 μl pipette tip available from MolecularBioProducts of San Diego, Calif. as Cat. No. 904-011. This particularpipette tip is especially preferred for applications where carryovercontamination is a concern, since it includes a filter (not shown)located at a position within an interior chamber 154 of the pipette tip70A-C, (see FIG. 18), which functions to block or impede the passage ofpotentially contaminating liquids or aerosols generated duringpipetting. Other acceptable pipette tips which can be modified asdescribed herein include the MβP® BioRobotix™ 1000 μl pipette tipavailable from Molecular BioProducts as Cat. No. 905-252 or 905-262.While the preferred number of lower ribs 151A-C, 152A-C is three, theprecise number selected should be determined, at least in part, by thetype of resin or combination of resins used to manufacture the pipettetip 70A-C, as well as the expected force needed to pierce a penetrablecap 20A-C or other surface material when puncturing is an intended useof the pipette tip 70A-C. Where a softer material is chosen formanufacturing the pipette tip 70A-C, or more force will be required topierce a surface, it may be desirable to increase the number of lowerribs 151A-C, 152A-C on the pipette tip 70A-C.

Another means by which to increase the rigidity of the pipette tip 70A-Cis to adjust the thickness or width of the lower ribs 151A-C, 152A-C. Ina preferred embodiment, the lower rib structure 151A which co-terminateswith the beveled tip 71A has a greater thickness and width than any ofthe other lower ribs 152A positioned on the pipette tip 70A. As shown inFIGS. 12 and 13, the larger of these preferred lower ribs 151Asubstantially forms a semi-circle in cross-section having a radius ofabout 0.020 inches (0.508 mm), whereas each of the smaller preferredlower ribs 152A, which also substantially form semi-circles incross-section, has a radius of about 0.012 inches (0.305 mm) in thispreferred embodiment. Of course, those skilled in the art will be ableto readily adjust the thicknesses and depths of the lower ribs 151A-C,152A-C by taking into consideration the properties of the resin selectedand the anticipated force needed to penetrate one or more pre-selectedsurface materials. And although the shape of the preferred lower ribs151A-C, 152A-C is substantially a solid semi-circle in cross-section,the lower ribs of the present invention may have either a solid orhollow core and can be constructed to include any one or a combinationof shapes (in cross-section), provided the shape or shapes of the lowerribs 151A-C, 152A-C do not significantly interfere with the penetrationor fluid-flow characteristics of the pipette tip 70A-C.

Although the preferred location of the lower ribs 151A-B, 152A-B is onthe outer surface 153 at the distal end of the pipette tip 70A-B,positioning the lower ribs on the inner surface 157 at the proximal endof the pipette tip 70C may have certain advantages. For instance,positioning the lower ribs 151C, 152C on the inner surface 157 of thepipette tip 70C could simplify the injection molding procedure by makingit easier and potentially less costly to prepare the molds.Additionally, positioning the lower ribs 151C, 152C on the inner surface157 may reduce the formation or extent of hanging drops on the bottomsurface (not shown) of the pipette tip 70C and reduce the adherence offluid to the outer surface 153 of the pipette tip by reducing thesurface area of the pipette tip which comes into contact with a fluid.In this particular configuration, the lower ribs 151A, 152A shown inFIGS. 10 and 11 could be positioned in a mirrored fashion on the insideof the conical section 166, as shown in FIG. 18, being careful to choosethicknesses for these internally positioned lower ribs, and adjustingthe size of an orifice 161 at the distal end of the pipette tip 70C, sothat the movement of fluids into or out of the pipette tip will not besubstantially impeded. One possible arrangement designed to avoidexcessive disruption of the flow of fluids into or out of the pipettetip 70C is shown in cross-section in FIG. 19. Determining appropriatedimensions for these internal, lower ribs 151C, 152C and the orifice 161size of the pipette tip 70C would require nothing more than routineexperimentation and would depend upon the particular application.

The preferred distal termini 162A, 163A of the lower ribs 151A, 152A, asshown in FIG. 12, are flush with and partially define the bottom surface158A at the distal end of the pipette tip 70A. Thus, when the pipettetip 70A has a beveled tip 71A, as depicted in FIGS. 10-12, the distalterminus 162A, 163A of each of the lower ribs 151A, 152A will share thesame angle as the beveled tip with respect to the longitudinal axis 72shown in FIG. 10. In the preferred pipette tip 70A, this angle is about30° to about 60°, more preferably about 35° to about 55°, and mostpreferably 45°±5°. However, it is not a requirement of the presentinvention that the distal termini 162A, 163A be flush with and partiallydefine the bottom surface 158A of the pipette tip 70A. For example,FIGS. 14 and 16 highlight an alternative configuration where the distalterminus 162B of the rib structure 151B tapers away from (rather thanforms) a point 155B of the beveled tip 156B, thus creating more of awedge-like shape to the point 155B of the pipette tip 70B. As FIGS.14-16 show, the lower ribs 151B, 152B can also be positioned so that thesurfaces of the distal termini 162B, 163B are not co-extensive with thebottom surface 158B at the distal end of the pipette tip 70B, but areinstead formed at a point longitudinally above the bottom surface 158B.(While only the smaller of the lower ribs 152B is actually depicted inthis manner in FIGS. 14-16, the distal terminus 162B of the larger ofthe lower ribs 151B could likewise be positioned above the bottomsurface 158B.) Decreasing the surface area of the bottom surface 158B,in a manner similar to that shown in FIG. 16, could be advantageous ifit is desirable to minimize fluid droplet formation at the distal end ofthe pipette tip 70B due to surface tension.

While the distal termini 163B of the lower ribs 152B shown in FIGS.14-16 are blunt-ended, alternative designs could be equally acceptable.As an example, the smaller lower ribs 152B could have a tapered shapesimilar to that shown in FIG. 14 for the larger lower rib structure151B. A tapered form of the smaller lower rib structure 152B mightterminate at the outer circumference 165B of the bottom surface 158Bshown in FIGS. 15 and 16 or at some point above the bottom surface 158B.Whatever shape or terminus location is selected for each lower ribstructure 151A-C, 152A-C, the primary considerations in most cases willbe the effect that the size, shape, number and positioning of the lowerribs 151A-C, 152A-C will have on air displacement from a collectiondevice 10 and/or the overall strength of the pipette tip 70A-C forpenetrating a pre-selected surface material.

The distance that the preferred lower ribs 151A-B, 152A-B extend awayfrom the distal termini 162A-B, 163A-B, which generally will be locatedat or near the bottom surface 158A-B of the pipette tip 70A-B, may varybetween lower ribs 151A-B, 152A-B on the same pipette tip 70A-B and maybe of any length, although preferred lengths are at least about 0.25inches (6.35 mm), at least about 0.5 inches (12.7 mm), and at leastabout 1.0 inch (25.4 mm). Where the distal termini 162A-B, 163A-B arelocated “near” the bottom surface 158A, 158B, the distance from an outerperimeter 165A, 165B at the distal end of the pipette tip 70A-B to eachdistal terminus 162A-B, 163A-B is no more than about 0.5 inches (12.7mm), and preferably no more than about 0.25 inches (6.35 mm) (thisdefinition of “near” is equally applicable to descriptions of the distaltermini (not shown) of lower ribs 151C, 152C positioned on the innersurface 157 of the conical section 166 and the continuous ribs 176described infra). In a preferred embodiment illustrated in FIGS. 10, 11,14 and 15, the pipette tip 70A-B forms a conical section 166 at thedistal end of the pipette tip 70A-B, and the lower ribs 151A-B, 152A-Bextend from or near the bottom surface 158A-B of the pipette tip 70A-Bto a point at the proximal end of the conical section 166, where theconical section 166 converges with a tubular section 167. (Opposingportions of the longitudinal wall defining the tubular section 167 neednot be parallel.) In this embodiment, the proximal terminus 168, 169 ofeach lower rib structure 151A-B, 152A-B tapers to a point where it meetsthe circumferential line 170 separating the conical section 166 from thetubular section 167. The lower ribs 151A-B, 152A-B may also extend froma point at or near the bottom surface 158A-B to any point on the tubularsection 167, even to a point at or near a top surface 173 at theproximal end of the pipette tip 70A-B (if no flange 172 is present) or,as shown in FIG. 20, a bottom surface 171 of the flange 172 at theproximal end of the pipette tip 70D.

By extending the lower ribs 151A, 152A to a point or points on thetubular section 167, (see, e.g., FIG. 20), or separately or exclusivelypositioning upper ribs 174 on the tubular section 167, (see FIGS. 14-18for examples of “separate” positioning and FIG. 21 for an example of“exclusive” positioning), benefits are expected to inhere when theintended use of the pipette tip 70B-E is to penetrate a surface materialassociated with a fluid-containing vessel 50. The most important ofthese benefits is the creation of air gaps or passageways 180, (see FIG.26, which illustrates penetration of a non-striated cap 20D), thatpermit at least a portion of the air displaced from a penetratedcollection device 10 to escape through openings created between thefluid transfer device and a penetrated surface material. Upon surfacepenetration, these passageways 180 form in areas adjacent contact points181 between the upper ribs 174 or continuous ribs 176 and the penetratedsurface material (e.g., a conical inner wall 33 for cap 20D of FIG. 26).By creating these passageways 180 during penetration, the upper ribs 174or continuous ribs 176 aid in preventing a high pressured movement ofair through openings in the penetrated surface material as the pipettetip 70B-E is being inserted into or withdrawn from a collection device10.

With fluid transfer devices having smaller diameters, such asfluid-transporting needles, air displacement by the fluid transferdevice entering a collection device 10 may be less of a concern.Notwithstanding, there may still be concerns about pressure differencesbetween the interior space of the collection device 10 and thesurrounding environment. When the air pressure inside of the collectiondevice 10 is sufficiently greater than the ambient air pressure, thenthere is a risk that at least some of the fluid material inside of thecollection device will escape through the opening created in apenetrated surface material when the fluid transfer device is withdrawnfrom the collection device. This is because the penetrated surfacematerial may form a seal around the entering fluid transfer device whichis largely broken when the fluid transfer device is completely withdrawnfrom the collection device 10, at which time fluid material in the formof an aerosol or bubbles may escape from the collection device as thetwo air pressures rapidly seek equilibrium. Moreover, because thepenetrated surface material may form a seal around the fluid transferdevice, a partial vacuum within the collection device 10 may be createdwhich could draw fluid material out of the fluid transfer device,thereby affecting pipetting accuracies and possibly leading to drippingof fluid material as the fluid transfer device is withdrawn from thecollection device. To minimize or eliminate these potential problems, itis important to provide a passageway for venting air from the collectiondevice 10 as the surface material is being penetrated by the fluidtransfer device and to maintain this passageway as the fluid transferdevice is withdrawn. This can be achieved by adding upper or continuousribs 174, 176 to at least some portion of the fluid transfer deviceexpected to be in contact with the surface material to be penetrated bythe fluid transfer device as it enters the collection device 10 toremove fluid material therefrom. In this way, small air gaps will becreated between the penetrated surface material and a portion of thefluid transfer device, thereby facilitating equilibrium between theinterior and exterior air pressures before the fluid transfer device isfully withdrawn from the collection device 10.

Where the upper ribs 174 are distinct from the lower ribs 151B, 152B, asshown in FIGS. 14-16, the upper ribs 174 are preferably aligned intandem with an equal number of lower ribs 151B, 152B positioned in alongitudinal orientation. The upper ribs 174 are preferably integrallymolded with the tubular section 167 using any well known injectionmolding process. While even one upper rib structure 174 could provide abeneficial air gap, at least three upper ribs 174 are preferred. Thereis, however, no set limit on the number of upper ribs 174 that may bepositioned on the tubular section 167. But where at least one purpose ofthe upper ribs 174 is to vent the interior chamber 175 of the collectiondevice 10, then the size, shape, number and orientation of the upperribs 174 should be chosen so that air gaps will be formed duringpipetting, thus facilitating adequate venting of displaced air and/orthe equilibration of air pressures inside and outside of the collectiondevice 10.

As with the lower ribs 151A-C, 152A-C, the upper ribs 174 may be of anyone or a combination of shapes, when viewed in cross-section, providedthe shape or shapes of the upper ribs 174 do not significantly interferewith the penetration characteristics of the pipette tip 70B-E whichincorporates them. The shapes of the upper ribs 174, when used inconjunction with lower ribs 151A-C, 152A-C, may be the same or differentthan the shapes of the lower ribs 151A-C, 152A-C. Preferably, thecross-sectional shape of each upper rib structure 174 is a squaremeasuring about 0.02 inches (0.508 mm) in width by about 0.02 inches(0.508 mm) in height (measuring from the outer surface 153 of thetubular section 167). The precise dimensions of the upper ribs 174 arenot critical, provided the upper ribs are capable of producing thedesired air gaps without significantly interfering with the penetrationcharacteristics of the pipette tip 70B-E.

As indicated above, the lower and upper ribs of the pipette tip 70D mayform continuous ribs 176, as shown in FIG. 20, thereby creating ribs 176which are unbroken between the conical and tubular sections 166, 167.Notwithstanding, the preferred pipette tip 70B incorporates distinctlower and upper ribs 151B, 152B, 174. In this preferred embodiment,which is depicted in FIGS. 14-16, the lower ribs 151B, 152B taper attheir proximal ends to form termini 168, 169, which terminate at thecircumferential line 170 delineating the conical and tubular sections166, 167. The upper ribs 174 in this preferred mode have blunt-endedtermini 177 at their distal ends which terminate at the circumferentialline 170, although the upper ribs 174 in another preferred embodimenttaper in a mirrored fashion to lower ribs 151B, 152B, terminating at thecircumferential line 170.

Another preferred fluid transfer device for use with the cap 20A-C ofthe present invention is illustrated in FIGS. 22-25. As shown, thepreferred embodiment of this fluid transfer device is a pipette tip 70Fwhich includes one or more grooves 178 which are preferably aligned in aspaced-apart, longitudinal orientation and are recessed from the outersurface 153 of the pipette tip. It was discovered that these grooves 178could be substituted for the upper ribs 174 depicted in FIGS. 14-19 and21 and used to channel air displaced from an interior chamber of acollection device 10 penetrated by the pipette tip 70F. In FIG. 27, itcan be seen that this channeling results from a passageway 182 formedbetween a groove 178 on an outer surface 153 of the pipette tip 70F,(see also FIGS. 22 and 23), and a penetrated surface of the collectiondevice 10. Thus, the boundaries of the passageway 182 are defined by thesurface of the groove 178 and that portion of the penetrated surfacewhich forms a canopy 183 over the groove 178. The penetrated surfaceshown in FIG. 27 is an outer surface 37 of a conical inner wall 33 of acap 20D which does not include striations 35. In all other respects,this cap 20D is identical to the cap 20A of FIGS. 2, 3 and 5.

In a preferred embodiment, the pipette tip 70F includes three grooves178 which are circumferentially spaced-apart at equal distances on theouter surface 153 of the pipette tip 70F. While the grooves 178 may beof any size or shape sufficient to facilitate the displacement of airfrom a penetrated collection device 10, the grooves 178 are preferablyrectangular in cross-section, (see FIG. 24), and have a width of 0.02in. (0.51 mm) and a depth of 0.01 in. (0.25 mm). To be fully effectivein facilitating the displacement of air from an enclosed chamber, thegrooves 178 should be positioned on at least a portion the outer surface153 of the pipette tip 70F where contact between the pipette tip 70F anda penetrated surface of the collection device 10 is expected. Therefore,the grooves 178 preferably extend at least one-third the length of afluid transfer device, more preferably at least one-half the length of afluid transfer device, and most preferably at least two-thirds thelength of a fluid transfer device. When the fluid transfer device isshaped to include a conical section 166 and a tubular section 167, asshown in FIGS. 22 and 23, at least one of the grooves 178 is preferablypositioned on at least a portion of the tubular section 167, and morepreferably extends the entire length of the tubular section 167. In aparticularly preferred embodiment, at least one of the grooves 178overlaps both the conical and the tubular sections 166, 167 of the fluidtransfer device.

Fluid transfer devices which include the grooves 178 of the presentinvention can also be used in conjunction with ribs extending from anouter surface of the fluid transfer device, such as those describedsupra and illustrated in FIGS. 10, 11, 17, 18 and 21. Particularlypreferred is the groove 178 and lower rib 151A, 152A combination of thepipette tip 70F shown in FIGS. 22 and 23. In this embodiment, lower ribs151A, 152A extend from the outer surface 153 of the conical section 166of the pipette tip 70F and have the same configuration and positioningas the lower ribs 151A, 152A of preferred pipette tip 70A which isdescribed above and depicted in FIGS. 10-13. At the approximate planarlocation where the proximal termini 169 of the lower ribs 152A begin totaper toward at the circumferential line 170 separating the conical andtubular sections 166, 167, distal termini 179 of the grooves 178 of thepipette tip 70F begin to taper toward their full recessed depth, whichis preferably reached by the point the grooves 178 intersect thecircumferential line 170. (In an alternative embodiment, the distaltermini 179 are not tapered but rather are blunt-ended.) This planaroverlap between the lower ribs 151A, 152A and the grooves 178 creates atransition region designed to ensure that air continues to be displacedfrom a collection device 10 as contact between the penetrated surfaceand the pipette tip 70F passes from the conical section 166 to thetubular section 167. Except for the flange 172 portion, the grooves 178of this preferred embodiment extend the entire length of the tubularsection 167.

To further facilitate penetration of the cap 20A-D, the fluid transferdevices 70A-F of the present invention preferably include a beveled tip71A-D, as shown in FIGS. 10, 12, 14, 16, 18 and 20-22. When a beveledtip 71A-D is employed, the distal end of the fluid transfer device 70A-F(e.g., fluid-transporting needle or pipette made of a resin) preferablyhas an angle of about 30° to about 60° with respect to the longitudinalaxis of the fluid transfer device 70A-F (the longitudinal axis for thefluid transfer devices of the present invention is the same as thelongitudinal axis 72 shown for the fluid transfer device 70 depicted inFIG. 7). Most preferably, the angle of the beveled tip 71A-D is about45°±5° with respect to the longitudinal axis of the fluid transferdevice 70A-E. However, a beveled tip of any angle that improves thepenetrability of a cap is desirable, provided the integrity of the fluidtransfer device is not compromised when the tip penetrates the cap,thereby affecting the ability of the fluid transfer device topredictably and reliably dispense or draw fluids.

In order to be useful, the fluid transfer devices of the presentinvention should be constructed so that their proximal ends can besecurely engaged by a probe associated with an automated or manuallyoperated fluid transfer apparatus. A fluid transfer apparatus is adevice which facilitates the movement of fluids into or out of a fluidtransfer device, such as a pipette tip. An example of an automated fluidtransfer apparatus is a GENESIS Series Robotic Sample Processoravailable from TECAN AG of Hombrechtikan, Switzerland, and an example ofa manually operated fluid transfer apparatus is the Pipet-Plus®Latch-Mode™ Pipette available from the Rainin Instrument Company ofEmeryville, Calif.

As an alternative to a fluid transfer device having ribs and/or groovesfor venting air displaced from an enclosed chamber of a collectiondevice, the present invention also contemplates a cap 20E featuring oneor more outwardly extending ribs 184 positioned on an inner surface 36of a conical inner wall 33, each rib 184 preferably having alongitudinal orientation. A preferred embodiment of this cap 20E isillustrated in FIGS. 28-30. As with the ribs of the fluid transferdevices 70A-F described above, the ribs 184 of this cap 20E are designedto form passageways 185 between the inner surface 36 of the conicalinner wall 33 of the cap and an outer surface 190 of a fluid transferdevice 70 as it is penetrating the cap, thereby permitting at least aportion of the air displaced from a vessel 50 associated with the cap toescape through these passageways 185. Upon surface penetration, thesepassageways 185 form in areas adjacent contact points 186 between theribs 184 of the conical inner wall 33 and the fluid transfer device 70,as depicted in FIG. 30. (To avoid cluttering FIGS. 28-30, those skilledin the art will appreciate that only some of the multiple striations 35,ribs 184, pie-shaped sections 26, passageways 185 and contact points 186which are clearly illustrated in these drawings are identified withreference numerals.) By creating these passageways 185 duringpenetration, the ribs 184 of the conical inner wall 33 help to prevent ahigh pressured movement of air through an opening in the conical innerwall, especially as the fluid transfer device is being removed from thecollection device. The ribs 184 of the cap 20E were also found to limitthe amount of frictional interference between the cap and the fluidtransfer device, making it easier to withdraw the fluid transfer devicefrom the penetrated cap.

While the ribs 184 may be incorporated into non-striated caps, caps 20Ehaving striations 35 are preferred. When the striations 35 are arrangedso that generally pie-shaped sections 26 are formed on a surface of theconical inner wall 33, a rib 184 having a longitudinal orientation ispreferably formed at the center of each pie-shaped section, asillustrated in FIG. 29. To limit the force required to penetrate a cap20E, the distal end of each rib 184 preferably terminates at a locationon the inner surface 36 of the conical inner wall 33 longitudinallyabove the apex 34, as shown in FIGS. 28 and 29. For applications inwhich the fluid transfer device is a pipette tip having a conicalsection 166 and a tubular section 167, such as the pipette tips 70A-Fshown in FIGS. 10-23, the ribs 184 are preferably arranged so thatcontact between the ribs 184 and the outer surface 153 of the conicalsection 166 is limited as the pipette tip initially pierces the apex 34.In this way, interference between the cap 20E and the pipette tip isminimized since it will be the tubular section 167 of the pipette tipwhich primarily makes contact with the ribs 184 of the cap.

In a particularly preferred embodiment, the approximate dimensions ofthe cap 20E depicted in FIGS. 28-30 are those specified infra in theExamples section. Additionally, the cap 20E of this preferred embodimentincludes eight ribs 184, each rib extending outwardly from theapproximate center of one of the pie-shaped sections 26 of the conicalinner wall 33 and having a longitudinal orientation. For this preferredembodiment, a proximal end of each rib 184 slopes outwardly from a pointabout 0.02 inches (0.508 mm) from the outer circumference 38 of theconical inner wall 33 at an angle of about 10° with respect to the innersurface 36 of the conical inner wall 33, for a total distance of about0.06 inches (1.52 mm). This proximal slope is built into the ribs 184 toprevent obstructing the downward movement of a misaligned fluid transferdevice which comes into contact with one of the ribs during a fluidtransfer operation. At the distal end of the slope, each rib 184 has agenerally parallel orientation with respect to the outer surface 37 ofthe conical inner wall and extends for a distance of about 0.09 inches(2.29 mm) before sloping inwardly toward the inner surface 36 of theconical inner wall 33 for a distance of about 0.015 inches (0.381 mm) atthe distal end of each rib 184. Based on this configuration, thegreatest thickness of these preferred ribs 184 is about 0.01 inches(0.254 mm), as measured outwardly at a right angle from the innersurface 36 of the conical inner wall 33. Moreover, each rib 184terminates at the distal end about 0.07 inches (1.78 mm) from the axisof symmetry 30, measuring at a right angle to the axis of symmetry. Thewidth of these preferred ribs 184 is about 0.015 inches (0.381 mm).

The present invention also contemplates ribs 184 which extend outwardlyfrom a penetrable surface of a cap which are of any size, shape ororientation sufficient to facilitate the formation of air passageways185 between the cap and a fluid transfer device but which do notsignificantly interfere with movement of the fluid transfer device intoor out of the penetrable cap. Accordingly, the ribs 184 may be elongatedstructures or they may be single protuberances or series ofprotuberances along a penetrable surface of the cap. The ribs 184 mayhave uniform orientations and be circumferentially spaced-apart at equaldistances from each other on a penetrable surface of the cap or they maybe arranged at different distances or in different orientations fromeach other. From this description, those skilled in the art will readilyappreciate ribs 184 of different shapes, dimensions and orientationswhich may be used to form air passageways 185 which will not createexcessive frictional forces between a penetrable cap and a fluidtransfer device.

To further minimize the frictional forces between a penetrable cap and afluid transfer device, it was advantageously discovered that apenetrable surface of the cap or an outer surface of the fluid transferdevice could be coated with a lubricant prior to piercing the cap.Lubricants contemplated by the present invention include, but are notlimited to, waxes (e.g., paraffin), oils (e.g., silicone oil) anddetergents (e.g., lithium lauryl sulfate). In a preferred mode, thelubricant is contained in a collection device and applied to apenetrable surface of the cap which is exposed to the interior of thecollection device by inverting the collection device one or more timesprior to penetration. As a consequence, lubricant from this cap surfacewill adhere to the outer surface of the fluid transfer device as itpenetrates the cap, thus minimizing frictional interference between thecap and the fluid transfer device when the fluid transfer device issubsequently withdrawn from the collection device. Moreover, when thelubricant is contained in the collection device, it is preferably acomponent of a specimen transport medium, such as lithium laurylsulfate. Detergent-containing transport mediums are well known in theart and would not have to be modified for this specific application.

Alternatively, the lubricant may be applied to an outer surface of thefluid transfer device or to a penetrable surface of the cap which isexposed to the exterior of the collection device. Lubricant may beapplied to the outer surface of the fluid transfer device by, forexample, dipping the fluid transfer device into a lubricant-containingtrough prior to penetrating the cap, where the trough is preferablysized to permit a majority of the outer surface of the fluid transferdevice to be coated with the lubricant. If this approach is followed,then, after submerging the fluid transfer device in thelubricant-containing trough, air should be expelled from the fluidtransfer device to remove any lubricant which may be obstructing thedistal orifice of the fluid transfer device prior to performing a fluidtransfer. With the cap, lubricant may be applied to the surface of thecap directly or by means of a lubricant-containing vesicle which can bepunctured by the fluid transfer device upon penetration of the cap. Inany case, the amount of lubricant applied to the cap should be limitedso that the distal orifice of the fluid transfer device does not becomeexcessively clogged with lubricant, thereby interfering with the fluidtransfer device's ability to draw fluids into its hollow body. Thoseskilled in the art will be able to make the appropriate adjustmentsbased on the configuration of the cap, the viscosity of the lubricantand the size of the fluid transfer device's distal orifice withouthaving to engage in undue experimentation.

Once a cap surface has been pierced, it is important to provide anenvironment that will allow for accurate aspirations of fluids,especially where the fluid will be employed in a volume sensitive assay.To this end, the applicants discovered that a two-step penetrationprocedure, which is preferably automated, resulted in more accuratefluid aspirations. Specifically, this procedure involves penetrating asurface of the cap at two distinct speeds. In a first step, the fluidtransfer device punctures the cap at a first speed, preferably in therange of about 15 to about 60 mm/s, followed by a second step, in whichthe fluid transfer device continues penetrating the cap at a secondspeed which is greater than the first speed and is preferably at leastabout 2 times, more preferably at least about 5 times and mostpreferably at least about 10 times the first speed. During the firststep, the distal end of the fluid transfer device preferably penetratesbeyond the punctured surface of the cap a distance of up to about 1 mm,2 mm, 3 mm, 5 mm, 10 mm, 15 mm or 20 mm. If the fluid transfer device isa plastic pipette tip, such as one of the pipette tips shown in FIGS.10-25, then it is preferred that some portion of the conical section 166be in contact with the penetrated surface of the cap after the firststep has completed.

Between the first and second steps, there is preferably a pause wherethe downward movement of the fluid transfer device is substantiallyarrested prior to initiating the second step. (The fluid transfer devicemay be withdrawn from the surface of the cap during this pause step.)This pause is preferably at least about 0.5 seconds in duration. It isduring this pause that the applicants speculate that air from theinterior of the collection device is vented, thereby minimizing vacuumformation as the fluid transfer device completes its penetration of thecollection device during the second step. The greater speed of thesecond step facilitates the opening of the penetrated surface, thushelping to form air passageways which promote air intake between thefluid transfer device and the penetrated surface of the cap. Incombination, the first and second steps aid in creating an environmentwithin the collection device which permits accurate aspirations offluids. And, assuming the applicants' venting theory is correct, thereshould also be some beneficial effect from carrying out the first andsecond steps at the same speed, provided a pause is introduced betweenthese two steps.

Another approach to facilitate the venting of air from within acollection and to achieve more accurate fluid aspirations is to use aconically-shaped pipette tip to penetrate a cap surface of thecollection device. With this approach, the pipette tip is inserted intoan interior chamber of the collection device a sufficient distance sothat a distal end of the pipette tip becomes at least partiallysubmerged in a fluid substance contained in the collection device. Thedistal end of the pipette tip is then partially or fully withdrawn fromthe fluid substance a sufficient distance to permit the formation orenlargement of one or more passageways between an outer surface of thepipette tip and the penetrated surface of the cap. (As used herein, a“passageway” is a space between an outer surface of a fluid transferdevice and a penetrated surface of a collection device (e.g., anassociated cap) which permits air from within the collection device topass into the surrounding environment.) In a preferred mode, the distalend of the pipette tip remains in contact with the fluid substance. Theformation or enlargement of the passageways may result when the surfacematerial of the cap is comprised of a less than fully resilientmaterial, such as HDPE, and the circumference of the pipette tipdecreases longitudinally from a proximal end to the distal end of thepipette tip. After these passageways are formed or enlarged, the pipettetip draws at least a portion of the fluid substance before the pipettetip is completely removed from the collection device. If the pipette tipis fully removed from the fluid substance when forming or enlarging thepassageways, then it will be necessary to reinsert the distal end of thepipette tip into the fluid substance prior to drawing fluid substancefrom the collection device. The steps of this procedure are preferablyautomated.

Returning to the description of the conical inner wall 33 depicted invarious embodiments in FIGS. 1-9, it should be pointed out that thenumber of striations 35 selected and the distance that those striations35 extend from start-points 31 at or near the apex 34 to the outercircumference 38 of the conical inner wall 33 should be sufficient tomaintain at least a portion of the generally wedge-shaped sections 26 ofthe conical inner wall 33 in an “open” configuration after the conicalinner wall 33 has been penetrated by a fluid transfer device and thefluid transfer device has been removed from the cap 20A-C. Asillustrated in FIG. 8, the wedge-shaped sections 26 of the conical innerwall 33 are in an “open” configuration provided that at least a portionof the tips 29 of the wedge-shaped sections 26 are not in physicalcontact with one another after the fluid transfer device has beenremoved from the cap 20A-C. (The conical inner wall 33 is deemed to bein the “open” configuration when at least two of the wedge-shapedsections have separated from one another after penetration of the cap20A-C by the fluid transfer device.) By maintaining the wedge-shapedsections 26 in an “open” configuration, frictional contact between thecap 20A-C and fluid transfer device is reduced and venting of air frominside of the collection device 10 is facilitated.

The distance that the striations 35 extend from the apex 34, orstart-points 31 near the apex 34, of the conical inner wall 33 to theouter circumference 38 of the conical inner wall 33 may be any distancesufficient to improve the penetrability of the conical inner wall 33 ascompared to an identical conical inner wall 33 having no striations 35.An improvement in penetrability is measured as a reduction in the forcerequired to penetrate the conical inner wall 33 of the cap 20A-C, asdescribed hereinabove. While it is not essential that all of thestriations 35 extend the same distance, it is preferred that eachstriation 35 extend radially outwardly at least about a quarter thedistance from the apex 34, or a start-point 31 near the apex 34, to theouter circumference 38 of the conical inner wall 33. In a more preferredmode, each striation 35 extends radially outwardly at least about halfthe distance from the apex 34, or start-points 31 near the apex 34, tothe outer circumference 38 of the conical inner wall 33. And in the mostpreferred embodiment of the present invention, each striation 35 extendsradially outwardly from the apex 34, or a start-point 31 near the apex34, to the outer circumference 38 of the conical inner wall 33.

Another factor to be considered in determining what distance thestriations 35 should extend from the apex 34 to the outer circumference38 of the conical inner wall 33 is the circumferential size of the fluidtransfer device. As the circumferential size of the fluid transferdevice increases, the distance that the striations 35 extend from theapex 34, or start-points 31 near the apex 34, to the outer circumference38 of the conical inner wall 33 will likewise need to increase in orderto improve penetration, allow for the formation of adequate airpassageways, and to minimize the frictional forces applied to fluidtransfer device by the conical inner wall 33 when the fluid transferdevice is entering or being withdrawn from the collection device 10.Increasing the number of striations 35 will also aid in reducing thefrictional forces applied by the conical inner wall 33.

Because the striations 35 may be formed as grooves, etchings or a seriesof perforations in the conical inner wall 33, the thicknesses of thestriations present in the conical inner wall—which may be the same ordifferent from one another—are less than the thicknesses of thesurrounding areas the conical inner wall. When determining the differentthicknesses of a conical inner wall 33, the cap 20A-C should first becooled at room temperature for a period of at least one hour afterforming, or cooled in tap water for at least 10 to 15 minutes, so thatthe resin can sufficiently harden. Four sections of the cap 20A-C, eachpreferably including a different striation 35 in cross-section, may thenbe cut at right angles to the striations 35 using an Xacto or utilityknife. With each of these sectional pieces of the conical inner wall 33of the cap 20A-C, a single measurement can be taken from each of thestriated and non-striated portions using any sensitive measuring means,such as calipers and/or video-based measuring instruments, in order todetermine the thicknesses between the inner and outer surfaces 36, 37 ofthe conical inner wall 33 in these portions. For the striated portions,the thickness measurements should be based on the smallestcross-sectional thickness between the inner and outer surfaces 36, 37.The thickness values thus obtained can be averaged to calculate theapproximate thicknesses of the striated and non-striated portions makingup the conical inner wall 35 of the cap 20A-C.

In a preferred embodiment, the thickness ratio, which is based on theratio of the average thickness of the non-striated portions of theconical inner wall 33 to the average thickness of the striations 35 inthe conical inner wall 33, is preferably in the range of about 5:1 toabout 1.25:1, more preferably in the range of about 7.5:1 to about 2:1,and most preferably in the range of about 10:1 to about 2.5:1. Theaverage thickness of the striations 35 of the conical inner wall 33 ispreferably in the range of about 0.002 inches (0.051 mm) to about 0.008inches (0.203 mm), and the average thickness of the non-striatedportions of the conical inner wall 33 is preferably in the range ofabout 0.01 inches (0.254 mm) to about 0.02 inches (0.508 mm). (Theindicated thicknesses for the striations are also the preferredthicknesses of the conical inner 33 when no striations 35 are included.)More preferably, the average thickness of the non-striated portions ofthe conical inner wall 33 is about 0.010 inches (0.254 mm) to about0.017 inches (0.432 mm); about 0.012 inches (0.305 mm) to about 0.015inches (0.381 mm); and about 0.013 inches (0.330 mm). At a minimum, thedifference in average thicknesses between the striations 35 and thenon-striated portions of the conical inner wall 33 should be such thatthe resistance encountered by the fluid transfer device as it passesthrough the conical inner wall 33 is less than it would be in theabsence of such striations 35, i.e, a conical inner wall 33 having asubstantially uniform thickness.

When the striations 35 include a series of perforations, theperforations are preferably sized to limit or prevent the passage offluid substance in the vessel 50 to the inner surface 36 of the conicalinner wall 33, where it could come into contact with a practitioner.This is particularly important where the fluid substance contains apotentially contaminating material (e.g., pathogenic organism). Tofurther ensure that no contaminating contact occurs between apractitioner and a fluid substance contained in the vessel 50 of thecollection device 10 when perforations constitute part or all of thestriations 35 in the conical inner wall 33, the seal 80 discussedhereinabove may be applied to the upper surface 24 of the annular topwall 22 (cap 20A-B) or to the annular top surface 48 (cap 20C) duringmanufacture so that the aperture leading to the conical inner wall 33remains completely enclosed.

Nonetheless, even when a seal 80 is employed, series of perforations donot constitute the preferred striations 35 of the present invention.This is especially the case where the collection device 10 will beshipped and potentially exposed to fluctuations in temperature andpressure which could result in fluid material leaking through theperforations, particularly where the collection device 10 is notexpected to remain upright during shipping. Additionally, fluid whichhas leaked through perforations present in the conical inner wall 33 tothe inner surface 36 could be absorbed by an optionally present wick 90,possibly causing the wick 90 to become saturated. Insertion of a fluidtransfer device through a wick 90 so affected may actually promoteaerosol formation and/or bubbling and, thus, the spread of potentialcontaminants. Accordingly, the use of series of perforations for thestriations 35 is not recommended except when it is certain thecollection device 10 will remain upright and will not be exposed toextreme changes in temperature and pressure.

As shown in FIGS. 5 and 6, the annular outer flange 40, 40A has an innersurface 41, 41A adapted to grip an upper portion 62, (see FIG. 1), ofthe outer surface 53 of the vessel 50, such that an essentiallyleak-proof seal between the cap 20A-C and the vessel 50 can established.More specifically, the essentially leak-proof seal may be createdbetween the lower surface 23 of the annular top wall 22, 22A of the cap20A-C and the upper surface 52 of the annular rim 51 of the vessel 50.Under normal handling conditions, this essentially leak-proof seal willprevent seepage of specimen from an interior chamber 175 of the vessel50 to an area of the outer surface 53 of the vessel which might becontacted by a practitioner during routine handling. Normal handlingconditions would not include the application of excessive and unusualforces (i.e., forces sufficient to puncture or crush a cap or vessel),as well as temperature and pressure fluctuations not typicallyexperienced in the handling and transport of collection devices.

The inner surface 41 of the annular outer flange 40 may be adapted, asdepicted in FIG. 5, to include a thread 42, which permits the cap 20A-Cto be screwed onto an upper portion 62 of the outer surface 53 of thevessel 50, (see FIG. 1), where the vessel has a mated thread 54. Themated threads 42, 54 facilitate an interlocking contact between thethread 42 of the cap 20A-B and the thread 54 of the vessel 50.Screw-type caps are well known in the art and skilled practitioners willreadily appreciate acceptable dimensions and means of manufacture.Ideally, the threads 42, 54 are integrally molded with the cap 20A-C andthe vessel 50, respectively.

Another adaptation to the inner surface 41A of the annular outer flange40A contemplated by the present invention is a snapping structure, asillustrated in FIG. 6. Here, the inner surface 41A of the annular outerflange 40A is adapted to include a rim 43 which can be snapped over amated rim 55 on the outer surface 53 of the upper portion 62 of thevessel 50 (see FIG. 1). These rims 43, 55 are preferably integrallymolded with the annular outer flange 40A of the cap 20C and the outersurface 53 of the vessel 50, respectively. In order to create thissnapping feature, the materials selected for constructing the cap 20Cand vessel 50 must be sufficiently resilient and the diameter of theinner portion 45 of the rim 43 on the cap must be sized to be less thanthe diameter of the outer portion 56 of the rim 55 on the vessel, sothat the inner portion 45 of the rim 43 on the cap, as defined by thecircumference of the inner portion 45 of the rim 43, can fit over theouter portion 56 of the rim 55 on the vessel, as defined by thecircumference of the outer portion 56 of the rim 55, without requiringthe application of a mechanical force. Moreover, the location of therims 43, 55 should be such that the lower portion 57 of the rim 55 onthe vessel 50 nests in an overlapping fashion on the upper portion 44 ofthe rim 43 of the cap 20C after the cap has been fitted onto the vessel.Moreover, when the rim 55 of the vessel 50 is nesting on the rim 43 ofthe cap 20C, an essentially leak-proof seal should be formed between thelower surface 23 of the annular top wall 22A of the cap and the uppersurface 52 of the annular rim 51 of the vessel.

Regardless of the approach adopted for physically and sealablyassociating the cap 20A-C and vessel 50, the essentially leak-proofnature of this arrangement can be further improved by including twosimple modifications to the cap, as illustrated in FIGS. 5 and 6. Thefirst modification would be to create an angled portion 47 on the innersurface 41, 41A of the annular outer flange 40, 40A at the point wherethe annular rim 51 of the vessel 50 and the annular outer flange 40, 40Amake contact. In this way, the frictional contact between the angledportion 47 of the inner surface 41, 41A and the annular rim 51 of thevessel 50 will create a more secure barrier to the passage of fluidsfrom within the vessel. (The space shown in these figures between thelower surface 23 of the annular top wall 22, 22A of the cap 20A-C andthe upper surface 52 of the rim 51 of the vessel 50 would benon-existent or less severe when the cap is securely fitted onto thevessel.) Additionally, the outer circumference 38 of the conical innerwall 33 can be modified to include an annular outer rim 39, (see FIG.5), or annular skirt 121, (see FIG. 6), which is designed to be infrictional contact with the inner surface 59 of the side wall 58 of thevessel 50 when the cap 20A-C and vessel are physically and sealablyassociated. Contact between the inner surface 59 of the side wall 58 andeither the annular outer rim 39 or an outer wall 122 of the annularskirt 121 should further impede the leaking of fluids from the vessel50.

An alternative to the annular outer flange 40, 40A described hereinabovewould be an annular flange (not shown) having an outer surface adaptedto grip the inner surface 59 of the side wall 58 within the open-ended,upper portion 62 of the vessel 50. Such an annular flange could beconstructed to frictionally fit within the upper portion 62 of thevessel 50 in a manner similar to that described above for gripping theouter surface 53 of the upper portion 62 of the vessel with the innersurface 41, 41 of the annular outer flange 40, 40A. In another form, theannular flange could be sized to fit snugly within the upper portion 62of the vessel 50 without the need to include a rim or thread on both theouter surface of the annular flange and the inner surface 59 of thevessel. In all other respects, this cap could be designed to include thefeatures described herein for the cap 20A-C, including a wick 90 and/orseal 80. It is also possible to remove the annular outer flange 40, 40Aaltogether, thereby converting the annular top wall 22 into an annularring (not specifically shown) having a lower surface which can beaffixed to the upper surface 52 of the annular rim 51 of the vessel 50using, for example, an adhesive (e.g., an inert glue).

To improve the seal formed between the annular rim 51 of the vessel 50and the lower surface 23 of the annular top wall 22, 22A of the cap20A-C when the vessel and cap are in fixed association, an annular seal(not shown) in the shape of an O-ring may be sized to fixedly nest onthe lower surface 23 of the annular top wall 22, 22A. The annular sealmay be an elastomeric material (e.g., neoprene) whose thickness ischosen so that snapping of the rim 43 of the cap 20C over the rim 55 ofthe vessel 50, or screwing the cap 20A-B onto the vessel 50 so thattheir respective threads 42, 54 are interlocking, is not prevented.

EXAMPLE

To determine the amount of force needed to penetrate a cap 20A-C of thepresent invention, a Universal Tension/Compression Tester (“CompressionTester”), Model No. TCD 200, and a force gauge, Model No. DFGS-50, wereobtained from John Chatillon & Sons, Inc. of Greensboro, N.C. Becausethe Compression Tester is an automated instrument, it allows for greaterreproducibility when determining the compression needed to penetrate acap that may not be possible following a purely manual approach.

All caps 20A-C used in this test were made of HDPE and had asubstantially uniform thickness of between about 0.0109 inches (0.277mm) and about 0.0140 inches (0.356 mm), except in the region of thestriations 35. The depth of the conical inner wall 33 of the cap 20A-Cwas about 0.29 inches (7.37 mm) as measured along the longitudinal axis30 of the cap from the plane of the outer circumference 38 of theconical inner wall 33 to the apex 34 of the same. The diameter of theouter circumference 38 of the conical inner wall 33 was about 0.565inches (14.35 mm). With all caps 20A-C tested, the conical inner wall 33had a single angle of about 35° or about 45° from the longitudinal axis30.

When caps 20A-C being tested included striations 35, the thickness ofthe conical inner wall 33 at the approximate center of each striation 35was in the range of about 0.0045 inches (0.114 mm) to about 0.0070inches (0.178 mm), where all striations 35 of any given cap were ofsubstantially the same thickness and had an approximate width of 0.015inches (0.381 mm). The total number of striations 35 for striated caps20A-C was always eight and the striations 35 were all formed on theinner surface 36 of the conical inner wall 33 during the injectionmolding process. Striations 35 of the caps 20A-C tested extended eitherfully or about half the distance from the apex 34 to the outercircumference 38 of the conical inner wall 33.

The caps 20A-C were threadingly secured to a vessel 50 measuringapproximately 13 mm×82 mm and made of polypropylene. In order tostabilize the collection devices 10 prior to penetration with the forcegauge, each collection device was secured in an aluminum block having ahole bored therein for receiving and stably holding the vessel 50component of the collection device. The precise method chosen forpositioning a collection device 10 under the force gauge is notcritical, provided the collection device is secured in a verticalposition under the force gauge, as judged by the longitudinal axis 30.

In evaluating the force required to penetrate a cap 20A-C, the vessel 50with attached cap was first centered under the force gauge with aGenesis series 1000 μl Tecan-Tip pipette tip force-fitted onto a 2 inch(50.8 mm) extension located at the base of the force gauge. The pipettetips were either blunt-ended or beveled with an angle of about 45° attheir distal ends. A cap 20A-C was considered to be centered when thepipette tip was located above the apex 34 of the conical inner wall 33of the cap. Absolute centering was not essential since the shape of theconical inner wall 33 of the cap 20A-C naturally directed the pipettetip to the apex 34 of the conical inner wall 33 of the cap. Since thepipette tip moved at a constant rate of 11.25 inches (285.75 mm)/minute,the initial height of the pipette tip above the cap 20A-C was notcritical, provided there was some clearance between the cap and thepipette tip. For testing purposes, however, the pipette tip wasgenerally positioned at least about 0.2 inches (5.08 mm) above the uppersurface 24, 24A of the annular top wall 22, 22A and permitted topenetrate up to 2.8 inches (71.12 mm) into the vessel 50, therebyavoiding actual contact with the inner surface 61 of the bottom wall 60of the vessel. The penetration force required was measured in poundsforce, and for all cap 20A-C tested the penetration force was less thanabout 6.5 pounds force (28.91 N). With fully-striated cap 20A-C andbeveled pipette tips, the penetration force was generally less thanabout 4.0 pounds force (17.79 N), and in some cases the penetrationforce required was about 3.6 pounds force (16.01 N) or less.

While the present invention has been described and shown in considerabledetail with reference to certain preferred embodiments, those skilled inthe art will readily appreciate other embodiments of the presentinvention. Accordingly, the present invention is deemed to include allmodifications and variations encompassed within the spirit and scope ofthe following appended claims.

1. A method for obtaining a fluid sample comprising: a) assembling acollection device comprising a cap and a fluid-holding vessel to containand isolate a specimen retrieval device adjacent a side wall of thevessel; b) entering the assembled collection device of step a) with afluid transfer device such that the specimen retrieval device does notsubstantially interfere with the movement of the fluid transfer deviceinto or out of the collection device; c) drawing at least a portion of afluid substance present in the collection device into the fluid transferdevice; and d) removing the fluid transfer device from the collectiondevice.
 2. The method of claim 1, wherein the cap and vessel includemated threads for fixedly associating the cap and vessel.
 3. The methodof claim 2, wherein the assembled collection device of step a) issubstantially leak-proof.
 4. The collection device of claim 1, whereinthe cap and vessel are each formed of an injection molded plastic. 5.The method of claim 1, wherein the specimen retrieval device is a swab.6. The method of claim 5, wherein the swab includes a score line tofacilitate snapping-off an upper portion of the swab above the scoreline.
 7. The method of claim 6 further comprising the steps ofsnapping-off and removing the upper portion of the swab.
 8. The methodof claim 1, wherein the fluid transfer device is a plastic pipette tip.9. The method of claim 8, wherein the penetrating step requires applyinga force of less than about 8 pounds force to a surface of the cap. 10.The method of claim 1, wherein the cap includes a plurality radiallyextending striations.
 11. The method of claim 1, wherein the fluidsubstance is suspected of containing a target nucleic acid.
 12. Themethod of claim 11 further comprising the step of amplifying a targetnucleic acid sequence contained in the target nucleic acid.
 13. Themethod of claim 12, wherein the amplifying step is performed using apolymerase chain reaction procedure.
 14. The method of claim 12, whereinthe amplifying step is performed using a transcription-mediatedamplification procedure.
 15. The method of claim 12 further comprisingthe step of detecting the presence of the target nucleic acid in thefluid substance.
 16. The method of claim 11 further comprising the stepof detecting the presence of the target nucleic acid in the fluidsubstance.
 17. The method of claim 1 further comprising the step ofinhibiting the release of an aerosol or bubbles from the collectiondevice.
 18. The method of claim 17, wherein the inhibiting step iscarried out using a wick contained within the cap or an overcap.
 19. Themethod of claim 18, wherein the wick is a pile fabric.
 20. The method ofclaim 12 further comprising the step of inhibiting the release of anaerosol or bubbles from the collection device.
 21. The method of claim20, wherein the inhibiting step is carried out using a wick containedwithin the cap or an overcap.
 22. The method of claim 21, wherein thecap or overcap includes a seal for retaining the wick within the cap orovercap.
 23. The method of claim 22, wherein the wick is a pile fabric.